In the cultivation of stem cells in vitro, components of animal origin and/or chemically undefined extracts like human platelet lysate are still used for the formulation of culture media because the quality of the most common basal media is unsuitable for cell culture in chemically defined environments. Experimental evidence shows that the better the qualitative and quantitative composition of the basal medium, the fewer additives of animal origin or chemically undefined components are needed to achieve successful cultivation of stem cells. The contribution deals with essential aspects concerning identity and quality of culture media components for pre-clinical and clinical research. It describes the development of physico-chemically defined micro-environments for cell cultures that modulate functions of the metabolism of human stem cells from adipose tissue (hASCs). The development process consists of two parts. The first phase involves the development of fully defined cellular microenvironments capable of satisfying the essential physiological needs of the cell. For this purpose, a detailed analysis of the physical (pH, osmotic pressure) and chemical requirements (energy sources, amino acids, vitamins, precursors of fatty and nucleic acids, inorganic salts, trace elements and chemical impurities) of the cells is carried out, which are then selected for the second phase. The second phase involves the elimination of complex, chemically undefined variables such as fetal calf serum or human platelet lysate, which are to be considered as critical components in cell culture media. The selection of a suitable basal medium and relative preparation technique allows the achievement of the goal. The results show that good knowledge of the variables that influence cellular growth processes has numerous advantages. The two-step strategy described above allows the identification of the essential physiological needs of the cells, thus favouring control and modulation of the main metabolic functions. This perspective becomes even more interesting as the omission of complex and expensive medium additives in clinical culture processes leads to a reduction of contamination risk and production costs.